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Bacillus subtilis subso natto for producing natto kinase and application thereof

A technology of Bacillus subtilis and nattokinase, applied in the biological field, can solve the problems of low cost of health care products, high cost of clinical standard drugs for nattokinase, and high cost of nattokinase

Inactive Publication Date: 2009-10-21
辽宁储阳生物科技开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, the industry has not carried out large-scale production of nattokinase. The first reason is that no bacterial strain suitable for large-scale fermentation of high-yield nattokinase has been found to reach the standard for industrialized production; the second is the cost of large-scale fermentation of nattokinase Relatively high, the cost of developing nattokinase clinical standard drugs is expensive, and the cost of developing corresponding health care products is correspondingly low

Method used

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  • Bacillus subtilis subso natto for producing natto kinase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Utilize bacillus subtilis natto subspecies LNUB236 to prepare nattokinase, its method is as follows:

[0045] 1) Strain activation: Bacillus subtilis subsp.natto LNUB236 was placed on LB slant medium, and cultured in a 37°C incubator for 24 hours;

[0046] The composition of LB slant medium is: tryptone 1g, sodium chloride 1g, yeast extract 0.5g, agar 1.5g, deionized water 100ml, pH 7.0;

[0047] 2) Seed liquid preparation: Pick the activated strains into two 500 ml Erlenmeyer flasks containing 200 ml LB liquid medium with an inoculation loop, and cultivate them at 37°C and 200 rpm for 18 hours;

[0048] LB liquid medium: 4 grams of tryptone, 4 grams of sodium chloride, 2 grams of yeast extract, 400 ml of deionized water, pH 7.0;

[0049] 3) Preparation of fermented liquid: Inoculate 2% seed liquid into the liquid comprehensive culture medium of fermenter, ferment temperature is 37°C, ferment time is 72 hours, pH is 7.0, ventilation rate is 0.5-1.0 cubic meters per hou...

Embodiment 2

[0055] A health product prepared by using nattokinase. The health product is proportioned by weight, natto freeze-dried powder:nattokinase:evening primrose oil=1:2.5:1.5.

[0056] Wherein, nattokinase is the nattokinase prepared in Example 1.

[0057] Preparation:

[0058] (1) Preparation of natto freeze-dried powder: soybeans were soaked in running water for 18 hours, then steamed in a pressure cooker at 121° C. for 90 minutes. After cooling, inoculate with an inoculum size of 40%, culture at 37°C for 1 day, and then culture at 4°C for 1 day, then mature natto, then add a small amount of distilled water to mash the mature natto with tissue crushed by a machine, and then freeze-dried in a vacuum to obtain freeze-dried natto powder.

[0059] (2) 15 grams of evening primrose oil is heated to remove oxygen therein and cooled to room temperature;

[0060] (3) Under aseptic conditions, 10 grams of natto freeze-dried powder, evening primrose oil and 25 grams of nattokinase were un...

Embodiment 3

[0062] A health product prepared by using nattokinase. The health product is proportioned by weight, natto freeze-dried powder:nattokinase:evening primrose oil=1:2.0:1.0.

[0063] Wherein, nattokinase is the nattokinase prepared in Example 1.

[0064] Preparation:

[0065] (1) Preparation of natto freeze-dried powder: soybeans were soaked in running water for 18 hours, then steamed in a pressure cooker at 121° C. for 90 minutes. After cooling, inoculate with an inoculum size of 40%, culture at 37°C for 1 day, and then culture at 4°C for 1 day, then mature natto, then add a small amount of distilled water to mash the mature natto with tissue crushed by machine, and then vacuum freeze-dried to obtain freeze-dried natto powder;

[0066] (2) 10 grams of evening primrose oil is heated to remove oxygen therein and cooled to room temperature;

[0067] (3) Under aseptic conditions, 10 grams of natto freeze-dried powder, evening primrose oil and 20 grams of nattokinase were uniform...

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Abstract

The invention relates to a bacillus subtilis subso natto for producing natto kinase and application thereof. The bacillus subtilis subso natto (bacillus subtilis subso natto LNUB236), CCTCC No.: M 208156. The bacillus subtilis subso natto is subject to activation of culture, seed liquid and fermentation liquor are prepared, and the natto kinase is extracted. The invention relates to application of the natto kinase to preparing a health product for preventing and treating cerebral thrombosis. Constituents of the health product are proportioned by weight: lyophilized natto powder : natto kinase : natto kinase : evening primrose oil=1:2.0-3.0:1.0-1.5. The bacillus subtilis subso natto LNUB236 has high natto kinase yield, the highest enzyme activity is 431.455IU / ml in the event of production in a 30l fermenter, and the prepared natto kinase-containing health product has the advantages of high enzyme activity, obvious effect and no side effect.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a bacillus subtilis natto subspecies capable of high-yielding nattokinase and an application thereof. [0002] A kind of Bacillus subtilis natto subspecies that produces nattokinase called in the present invention is named: Bacillus subtilis subsp. Preserved by the depository center, the number is CCTCC NO: M 208156. Background technique: [0003] Thromboembolic disease is one of the leading causes of death and is the second most common disease after cancer today. At present, dissolving thrombus is an important means to treat this kind of diseases. The drugs used in clinical treatment today include streptokinase, urokinase, tissue-type plasminogen activator, single-chain urokinase-type plasminogen activator, Chemically modified plasmin-streptokinase activator complex, etc. These drugs all activate plasminogen in the blood to degrade fibrin and act indirectly. The cli...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/56A61K38/48A61K9/48A61P7/02A61P9/10C12R1/125
Inventor 刘宏生寇巍李秀瑜
Owner 辽宁储阳生物科技开发有限公司
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